Monocyte Chemoattractant Protein-Induced Protein 1 Overexpression Modulates Transcriptome, Including MicroRNA, in Human Neuroblastoma Cells
Monocyte Chemoattractant Protein-Induced Protein 1 Overexpression Modulates Transcriptome, Including MicroRNA, in Human Neuroblastoma Cells
复制标题
DOI：
10.1002/jcb.25354
复制
发表时间：
2016-03-01
影响因子：
4
通讯作者：
Rokita, Hanna
中科院分区：
文献类型：
--
作者：
Boratyn, Elzbieta;Nowak, Iwona;Rokita, Hanna
最近发现的MCPIP 1（monocyte chemoattractant protein-induced protein 1）是由MCPIP 1（ZC 3 H12 A）基因编码的一种多结构域蛋白，被描述为一种新的分化因子、核糖核酸酶和去泛素化支持因子。然而，它在癌症中的作用却很少被认识到。我们最近对微阵列数据的分析显示，在儿童最常见的颅外实体瘤--原发性神经母细胞瘤中缺乏MCPIP 1转录本的表达。此外，BE（2）-C细胞中MCPIP 1基因的强制表达导致神经母细胞瘤增殖和活力显着降低。本研究的目的是利用表达DNA微阵列和microRNA微阵列进一步研究MCPIP 1在神经母细胞瘤中的作用。BE（2）-C细胞的瞬时转染用于过表达野生型MCPIP 1（MCPIP 1-wt）或其RNase缺陷型突变体（MCPIP 1-DPIN）。我们分析了转录组的变化，接下来，我们使用qRT-PCR来验证响应于MCPIP 1过表达的选定基因的mRNA水平。此外，确定了一些选定基因的蛋白质水平。由SLC 44 A1基因编码的胆碱转运蛋白CTL 1在特定的mRNA和蛋白水平上被显著抑制，最重要的是，这转化为MCPIP 1过表达细胞中胆碱转运的减少。然后，我们发现microRNA-3613- 3 p是过表达野生型MCPIP 1的细胞库中最大的改变。接下来，我们分析了miR-3613- 3 p的预测靶点，并使用qRT-PCR和western blot对其进行验证。这些结果表明，miR-3613- 3 p的表达可能是由MCPIP 1通过切割其前体形式来调节的。J.细胞。117：694-707，2016. (C)2015 Wiley Periodicals，Inc.
The recently discovered MCPIP1 (monocyte chemoattractant protein-induced protein 1), a multidomain protein encoded by the MCPIP1 (ZC3H12A) gene, has been described as a new differentiation factor, a ribonuclease, and a deubiquitination-supporting factor. However, its role in cancer is poorly recognized. Our recent analysis of microarrays data showed a lack of expression of the MCPIP1 transcript in primary neuroblastoma, the most common extracranial solid tumor in children. Additionally, enforced expression of the MCPIP1 gene in BE(2)-C cells caused a significant decrease in neuroblastoma proliferation and viability. Aim of the present study was to further investigate the role of MCPIP1 in neuroblastoma, using expression DNA microarrays and microRNA microarrays. Transient transfections of BE(2)-C cells were used for overexpression of either wild type of MCPIP1 (MCPIP1-wt) or its RN-ase defective mutant (MCPIP1-DPIN). We have analyzed changes of transcriptome and next, we have used qRT-PCR to verify mRNA levels of selected genes responding to MCPIP1 overexpression. Additionally, protein levels were determined for some of the selected genes. The choline transporter, CTL1, encoded by the SLC44A1 gene, was significantly repressed at the specific mRNA and protein levels and most importantly this translated into a decreased choline transport in MCPIP1-overexpressing cells. Then, we have found microRNA-3613-3p as the mostly altered in the pools of cells overexpressing the wild type MCPIP1. Next, we analyzed the predicted targets of the miR-3613-3p and validated them using qRT-PCR and western blot. These results indicate that the expression of miR-3613-3p might be regulated by MCPIP1 by cleavage of its precursor form. J. Cell. Biochem. 117: 694-707, 2016. (C) 2015 Wiley Periodicals, Inc.